Object To investigate the in vivo antioxidant effect of the extract from Rosmarinus officinalis L. and its active substances. Methods The contents of MDA, the activites of SOD and GSH Px in serum, heart, liver, brain and skeleton muscle were determined in oxidative stress mouse model caused by exercise. Results It was found that in serum, liver, heart and skeleton muscle except the brain, the contents of MDA were decreased and the activities of SOD and GSH Px were increased by 250 mg/kg and 500 mg/kg of total phenolic diterpenes (TPD) extract taken. Conclusion The results showed that R. officinalis has prominent antioxidant effect in exercise mice and the active constituents may be phenolic diterpenes.

Objective To construct myeloid-specific Spi1 gene knockout mice and analyze their genotypes,so as to provide animal model basis for the study of pathological mechanism of diseases and drug targets.Methods Ac-cording to the principle of CRISPR/Cas9 technology and Cre/LoxP system,sgRNA and Donor vectors were de-signed and constructed.The transcript of Exon 2(Exon 2)was used as the knockout region,and Loxp elements were placed on both sides of Exon 2.Cas9 protein,sgRNA and Donor vector were mixed and microinjected into the fertilized eggs of C57BL/6J mice,the fertilized eggs were transplanted into the uterus of C57BL/6J pregnant female mice,and F0 generation was obtained after 19～20 days.Positive F0 mice were mated with C57BL/6J mice to ob-tain stable F1 Spi1flox/+mice.Spi1flox/+mice of F1 generation were selfed to obtain Spi1flox/flox mice.Spi1flox/flox mated with Lyz2-Cre+mice to obtain Spi1flox/+/Lyz2-Cre+mice,and then mated with Spi1flox/flox,the Spi1flox/flox/Lyz2-Cre+mice were myeloid-specific Spi1 gene knockout(KO)mice.Spi1flox/flox/Lyz2-cre-mice were used as wild-type(WT)mice.DNA of WT and KO mice was extracted,and the genotypes were identified by agarose gel electro-phoresis after PCR amplification.Western blot was used to detect the expression of spleen focus forming virus provi-ral integration oncogene,Spi-1/purine rich box-1(PU.1)in immune cells of WT and KO mice.Results The results of PCR identification showed that the genotype of mice with only 220 bp amplified by flox primer was Spi1flox/flox homozygote,and the genotype of mice with 700 bp amplified by Lyz2-Cre primer was Lyz2-Cre+.Western blot showed that compared with WT group,the protein PU.1 was not expressed in bone marrow-derived macropha-ges(BMDMs)and peritoneal macrophages(PM)in KO group(P<0.01).There was no significant difference of statistics in the expression level of PU.1 in T cells between KO mice and WT mice.The results of PCR and West-ern blot showed that myeloid-specific Spi1 KO mice were successfully constructed.Conclusion The myeloid-spe-cific Spi1 gene KO mice are successfully constructed and identified,which provides animal model basis for further revealing the potential mechanism of PU.1 inimmune regulation.

Objective To construct Csf1r-CreERT2 R26REYFP reporter gene mice and assess the efficacy of Csf1r-CreERT2-mediated enhancement of CSF1R in CD45+cells labeled with yellow fluorescein protein EYFP.Methods Csf1r-CreERT2 mice were crossbred with R26REYFP homozygous mice,and Csf1r-CreERT2R26REYFP mice were identified through PCR and Western Blot analyses.Flow cytometry was employed to evaluate CSF1R tag-efficiency in CD45+cells across different mouse tissues following tamoxifen induction.Results Csf1r-CreERT2 R26REYFP reporter gene mice were acquired.In addition,it was found that Csf1r-CreERT2-mediated EYFP could effectively mark CSF1R in various tissues of mice and CD45+cells in different locations.Compared to the R26REYF P group,the highest labeling efficiency was observed in the brain tissue(P＜0.001),the lowest in the thymus tissue(P＜0.05),and no sig-nificant difference was observed in the spleen tissue.Conclusion Adult Csf1r-CreERT2 mice and R26REYFP mice are effective ways to obtain Csf1r-CreERT2 R26REYFP induced conditional fluorescence mice.Csf1r-CreERT2 can mediate EYFP to effectively trace CSF1R in CD45+cells in different parts of mice.

Objective:To explore effects of application of inadventent perioperative hypothermia (IPH) cluster nursing in surgical patients, and knowledge, belief and practice of IPH protection among Operating Room nurses.Methods:Using the convenient sampling method, a total of 2 166 surgical patients from Xiangyang Central Hospital from October 2018 to October 2020 were selected as the research objects. A total of 1 123 patients who underwent surgery from October 2018 to October 2019 were set as the control group, and 1 043 patients who underwent surgery from November 2019 to October 2020 were set as the observation group. Fifteen Operating Room nurses were included in the control group and observation group respectively. The nurses in the control group and in the observation group respectively adopted routine heat preservation measures and cluster nursing measures to carry out temperature nursing of surgical patients. The body temperature of patients in the two groups before and after operation were compared, as well as knowledge, attitude and practice questionaire of IPH protection of nurses of the two groups before and after the intervention.Results:The postoperative body temperature of the observation group was (36.51±0.79) ℃, which was higher than that of the control group (34.27±0.76) ℃, and the difference was statistically significant ( P<0.05) . After the intervention, the total score and section scores of nurses in the observation group on knowledge, attitude and practice questionaire of IPH protection were higher than that of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:IPH clustered nursing can reduce the occurrence of IPH in surgical patients and improve the knowledge, belief and practice level of IPH protection among Operating Room nurses.

Objective To breed and identify the T lymphocyte-conditional Spi1 knockout mice for the further in-vestgation of the specific role of Spi1-encoded protein PU.1.Methods The Lck-Cre mice were mated with Spi1flox/flox mice to obtain Lck-Cre×Spi1flox/flox mice(T lymphocyte-specific Spi1 knockout mice),and the genotype was determined by polymerase chain reaction(PCR)and agarose gel electrophoresis.Magnetic beads were used to sort out the splenic T lymphocytes,and the knockdown efficiency of PU.1 in T cells was detected by Western blot,quantitative real-time PCR(qPCR)and flow cytometry.Results The Lck-Cre×Spi1flox/flox mouse genotype was stably inherited.Compared with Spi1flox/flox mice,the expression level of PU.1 was significantly reduced in splenic T cells of Lck-Cre×Spi1flox/flox mice.Conclusion In this study,the T lymphocyte-specific Spi1 knockout mice was successfully constructed by applying Cre/LoxP system and CRISPR/Cas9 technology,which provided a reliable an-imal model for the subsequent experiments of the specific role of PU.1 in T cell-related diseases.

Objective To analyze the spatiotemporal distribution characteristics of the syphilis epidemic in Gansu Province from 2005 to 2021, and to provide a reference for the prevention and control of the syphilis epidemic in Gansu Province. Methods ArcGIS 10.7 was used to map the annual incidence of syphilis in Gansu Province from 2005 to 2021, spatial autocorrelation analysis and local autocorrelation analysis were performed, and SaTScan 10.0.2 software was used for spatiotemporal scanning analysis. Results The global autocorrelation results showed that the annual incidence of syphilis in 2005-2021 was >0, Z＞1.96, and the P< 0.0001, showing a spatial clustering distribution, and the local autocorrelation results showed that there was one spatially similar high-high aggregation area and two spatially similar low-low aggregation areas in Gansu Province, and the hot spot analysis showed that there were 9 negative hotspot areas and 2 positive hotspot areas in the syphilis epidemic in Gansu Province. Spatiotemporal scanning analysis detected two high concentration areas, mainly concentrated in Gannan Tibetan Autonomous Prefecture. Conclusion Syphilis in Gansu Province has regional differences in space, and high-high accumulation areas in Gannan Tibetan Autonomous Prefecture persist, and targeted prevention and control strategies should be specified according to temporal and spatial characteristics.

OBJECTIVETo investigate the role of CD147 gene on the proliferation and infiltration of a human monocytic leukemic cell line SHI-1.

METHODSThe expression of CD147 in SHI-1 cells was knockdowned by the lentiviral vector. The expressions of CD147, MMP-2 and MMP-9 were detected by semiquantitative RT-PCR. The protein of CD147 was detected by Western blotting. The capabilities of proliferation and infiltration of SHI-1 cell were examined by MTT and trans- matrigel invasion assay co-cultured with leukemia BMSC in vitro. SHI-1 cells were inoculated subcutaneously or via tail vein into nude mice to investigate its growth and infiltrative ability in vivo.

RESULTSThe mRNA and protein of CD147 in SHI-1/CD147i cells decreased by 85% and 91%, respectively after the SHI-1 cells were infected by the lentivirus containing the CD147 siRNA. The proliferation capability of SHI-1/CD147i cells significantly decreased than those of SHI-1 and SHI-1/NC cells. The mRNA expressions of MMP-2, MMP-9 in SHI-1/CD147i cells were significantly lower than those in SHI-1/NC and SHI-1 cells. The SHI-1/CD147i cells showed significantly lower invasion rate than SHI-1 cells and SHI-1/NC cells when co-cultured with BMSCs. The neoplasms formed by SHI-1/CD147i cells in the subcutaneous of mice were significantly smaller than of the neoplasms formed by SHI-1 and SHI-1/NC cells. In nude mice inoculated via caudal vein with SHI/CD147i cells, mice demonstrated longer survival and moderate infiltration characteristic than those inoculated with SHI and SHI-1/NC cells.

CONCLUSIONCD147 might play important roles in the proliferation and infiltration of leukemia cells. CD147 should be a potential target for the treatment of acute leukemia.

Animals ; Basigin ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Gene Knockdown Techniques ; Humans ; Lentivirus ; genetics ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics

Objective: To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected． Methods : CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) ．CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro．The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression，and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ，and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry. Results : CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells，flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P＜0. 01) ． Conclusion In this experiment，CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ，and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.